Figure 3.

In Vitro dsRNA Processing by Dcl5 Using Templates Containing Paramecium MAC and IES Sequences

(A) dsRNA templates used for in vitro assays with Dcl5 are shown schematically: a 500 nt long fragment of ND7 coding sequence (black line) and two templates made of three concatenated IESs (blue, purple, and orange).

(B) 15% polyacrylamide-urea gel with small RNAs 5′-labeled with ³²P. M is the small RNA ladder. (+) and (−) indicate reactions performed with or without the enzyme. Dashed boxes indicate the bands corresponding to the most abundant products after sRNA sequencing.

(C) Size distribution graphs of the cleavage products for Dcl5 with Concatemer and Concatemer Longer templates.

(D and E) Enrichment graphs for the 5′ (D) and 3′ (E) ends of Dcl5 cleavage products.